摘要
克隆中国对虾(Fenneropenaeus chinensis)胰蛋白酶基因,参考多种生物的胰蛋白酶基因序列设计出1对简并PCR引物。利用PCR技术从中国对虾基因组DNA中扩增出1条DNA带,经PCR产物回收、纯化,连接到载体质粒,转化细菌,蓝百斑筛选等操作获得了1个阳性克隆。序列分析表明,该序列包含1个内含子和2个不完整的外显子,与已报道的凡纳滨对虾(Litopenaeus vannamei)胰蛋白酶基因Ⅲ有较高的相似度(identity=79.2%)。综合多种分析,可以推定该序列为对虾的胰蛋白酶基因序列。系统进化分析表明该基因属于胰蛋白酶第三家族。
Trypsin is an important enzyme in the digestive gland of animals. In this paper, we report the cloning and characterization of a gene fragments encoding trypsin of Chinese Prawn （Fenneropenaeus chinensis）. According to the conserved domain of some trypsin genes reported in the GenBank, a pair of degeneration primers were designed and synthesized. Polymerase chain reaction（PCR） was performed using genomic DNA of Chinese Prawn as template. A trypsin gene clones was obtained after a series of manipulation, purification of PCR production, ligation and transformation of recombinant plasmids, blue-white selection etc. DNA sequence analysis showed that the sequence was composed of one intron and two incomplete extrons. The sequence had 79.2 % identity with trypsinogen Ⅲ of Pacific White Shrimp（Litopenaeus vannamei） and that the amino acid sequence deduced from the sequence had 92.3 96 identity with the one of Pacific White Shrimp. Therefore, the genomic DNA fragments can prove to be the incomplete trypsinogen of Fenneropenaeus chinensis （ try Fc ）. The phylogenetic tree shows that try Fc belongs to the third family of trpsinogen.
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第6期1017-1020,1052,共5页
Periodical of Ocean University of China
基金
中国海洋大学引进人才科研启动经费项目资助
